Aseptic Seedling Production by In Vitro Seed Culture in Salvia fruticosa Mill.

Year 2022, Volume: 12 Issue: 1, 11 - 19, 30.06.2022

Emel Yılmaz Gökdoğan , Müge Etik Betül Bürün

https://doi.org/10.54370/ordubtd.1028243

Abstract

The genus Salvia, which is a member of the Lamiaceae family, is a medicinally and aromatically important genus. The species Salvia fruticosa Mill., which grows in our country and is called Anatolian sage, causes both the increase of its economic value and its trade by collecting from nature because of containing the essential oil content of this species is higher than the other species. In this study, the germination of Salvia fruticosa seeds in 1/10 Murashige-Skoog (MS) (1962) nutrient medium was investigated obtaining aseptic seedlings for use in in vitro cultures. The seeds before culture were soaked in 70% ethanol for 2 minutes and then they were applied with NaOCl (chlorine), H2O2 (oxidant) and H2SO4 (acid). These seeds were divided into three groups: no cold treatment was applied to the 1st and 2nd group seeds, cold application was made to the 3rd group. While the first group seeds were taken to photoperiodic conditions after one week of darkness, the seeds in second and third groups were kept in photoperiodic conditions. The highest germination percentage was 50%, and it was obtained from the culture of seeds superficially sterilized with 30-31% H2O2 for 3 minutes in photoperiodic conditions after dark. In the germination test (control) of seeds on filter paper, germination was 48% under the same condition. The infection rate under the fotoperiodic condition was observed to be low in cold applied seeds. As a result, the procedure using H2O2, which gives the highest germination percentage (50%), is applicable for the production of sterile seedlings that can be used as explants in in vitro cultures of Salvia fruticosa for different purposes.

Keywords

anatolian sage, hydrogen peroxide, In vitro germination, sodium hypochlorite, sulfuric acid

Salvia fruticosa Mill.’da İn Vitro Tohum Kültürü ile Aseptik Fide Üretimi

Abstract

Lamiaceae familyası üyelerinden olan Salvia cinsi tıbbi ve aromatik olarak önemli bir cinstir. Ülkemizde yetişen ve Anadolu adaçayı olarak isimlendirilen Salvia fruticosa Mill. türünün uçucu yağ içeriğinin diğer türlere göre daha yüksek olması, doğadan toplanarak ticaretinin yapılmasına ve ekonomik değerinin artmasına neden olmaktadır. Bu çalışmada Salvia fruticosa tohumlarının 1/10 Murashige-Skoog (MS) (1962) besin ortamında çimlendirilmesi ile in vitro kültürlerde yararlanmak üzere aseptik fide elde edilmesi araştırılmıştır. Kültür öncesi tohumlar %70 etil alkole 2 dakika batırılmış ve takiben NaOCl (klor), H2O2 (oksidan) ve H2SO4 (asit) ile muamele edilmiştir. Bu tohumlar üç gruba ayrılmış: 1. ve 2. grup tohumlara soğuk uygulaması yapılmamış, 3. gruba soğuk uygulaması yapılmıştır. 1. grup tohumlar 1 hafta karanlığı takiben fotoperiyodik koşullara alınırken; 2. ve 3. gruptaki tohumlar fotoperiyodik koşulda tutulmuşlardır. En yüksek çimlenme %50’dir ve 3 dakika %30-31 H2O2 ile yüzeysel sterilizasyonu yapılmış tohumların karanlığı takiben fotoperiyodik koşuldaki kültüründen elde edilmiştir. Aynı koşuldaki tohumların filtre kâğıdı üzerindeki çimlenme testinde (kontrol) ise çimlenme %48 olmuştur. Soğuk uygulama yapılan ve fotoperiyodik koşulda tutulan tohumlarda ise enfeksiyon, düşük gözlenmiştir. Sonuç olarak, Salvia fruticosa’nın farklı amaca yönelik in vitro kültürlerinde eksplant olarak kullanılabilecek steril fidelerin üretimi için en yüksek çimlenme yüzdesini (%50) veren H2O2’nin kullanıldığı prosedür uygulanabilir özelliktedir.

Keywords

anadolu adaçayı, hidrojen peroksit, in vitro çimlenme, sodyum hipoklorit, sülfürik asit

References

• Arıkat, N. A., Jawad, F. M., Karam, N. S. ve Shibli, R. A. (2004). Micropropagation and accumulation of essential oils in wild sage (Salvia fruticosa Mill.). Scientia Horticulturae, 100, 193-202. http://doi.org/10.1016/j.scienta.2003.07.006
• Bakhsh, A., Anayol, E., Sancak, C. ve Özcan, S. 2016. An efficient and cost effective sterilizing method with least microbial contamination and maximum germination ratio for in vitro cotton (Gossypium hirsutum L.) culture. The Journal of Animal & Plant Sciences, 26(3), 868-873. https://www.researchgate.net/publication/303696595
• Barampuram, S., Allen, G. ve Krasnyanski, S. (2014). Effect of various sterilization procedures on the in vitro germination of cotton seeds. Plant Cell, Tissue and Organ Culture, 118, 179-185. http://doi.org/10.1007/s11240-014-0472-x
• Baran-Ayaz, Ö., Surgun, Y. ve Bürün, B. (2015). Endemik Anadolu sığlası (Liquidambar orientalis Mill.)’nda tohum sterilizasyonu ve çimlenme. Anadolu Doğa Bilimleri Dergisi, 6(Özel sayı 2), 82-86. http://plant-materials.nrcs.usda.gov
• Bayraktar, Ö. V., Öztürk, G. ve Arslan, D. (2017). Türkiye’de bazı tıbbi ve aromatik bitkilerin üretimi ve pazarlanmasındaki gelişmelerin değerlendirilmesi. Tarla Bitkileri Merkez Araştırma Enstitüsü Dergisi, 26 (2), 216-229. http://doi.org/10.21566/tarbitderg.369928
• Bayram, E., Kırıcı, S., Tansı, S., Yılmaz, G., Arabacı, O., Kızıl, S. ve Telci İ. (2010, Ocak, 11-15). Tıbbi ve aromatik bitkiler üretiminin arttırılması olanakları. Ziraat Mühendisliği VII. Teknik Kongresi, Ankara, Türkiye.
• Elena, T. M., Camen, D., Moatăr, M., Petolescu, C., Boldea, M., Iordanescu, O. ve Sala, F. (2019). Research regarding the influence of the preparing methods on seed germination on Salvia officinalis. Journal of Horticulture, Forestry and Biotechnology, 23(2), 53-57. https://journal-hfb.usab-tm.ro
• Ghanbar, T., Hosseini, B. ve Jabbarzadeh, Z. (2012). Improving Salvia sclarea L. seed germination under in vitro condition. International Journal of Agriculture: Research and Review, 2(S), 1051-1058. http://facultystaff.urmia.ac.ir
• Ghanbar, T., Hosseini, B., Jabbarzadeh, Z., Farokhzad, A. ve Sharafi, A. (2016). High-frequency in vitro direct shoots regeneration from axillary nodal and shoot tip explants of clarysage (Salvia sclarea L.). Bulgarian Journal of Agricultural Science, 22(1), 73-78. https://www.researchgate.net/publication/296675191
• Grzegorczyk, I., Bilickowski, I., Mikiciuk-Olasık, E. ve Wysokińska, H. (2005). In vitro cultures of Salvia officinalis L. as a source of antioxidant compounds. Acta Societatis Botanicorum Poloniae, 74(1), 17-21. https://doi.org/10.5586/asbp.2005.003
• Huii, L., Guoping Z., Guozheng, S., Songlin, R. ve Qiaojuan, F. (2012). Callus induction and plant regeneration from mature seeds of Salvia splendens. International Journal of Agriculture and Biology, 14(3), 445-449. https://www.fspublishers.org/published_papers/73876_..pdf
• Ioja-Boldura, O.M., Radu, F., Popescu, S. ve Borozan, A. (2010). Regeneration, micropropagation, callus cultures and somatic embryogenesis of common sage (Salvia officinalis L.). Bulletin UASVM Horticulture, 67(1), 308-313. DOI:10.15835/BUASVMCN-HORT:5732
• ISTA (1993). International Rules For Seed Testing, International Seed Testing Association. Seed Sci Technol 21, p. 289, Zürich, Switzerland.
• İpek, A. ve Gürbüz, B. (2012). Türkiye florasında bulunan Salvia türleri ve tehlike durumları. Tarla Bitkileri Merkez Araştırma Enstitüsü Dergisi, 19(1-2), 30-35. https://dergipark.org.tr/tr/download/article-file/118448
• Javaid, M.M., Florentine, S., Ali, H.H. ve Weller, S. (2018). Effect of environmental factors on the germination and emergence of Salvia verbenaca L. cultivars (verbenaca and vernalis): An invasive species in semi-arid and arid rangeland regions. PLOS ONE, 13(3), 1-20. https://doi.org/10.1371/journal.pone.0194319
• Kaczperski, M. P. ve Carlson, W.H. (1989). A Commercial Growers Guide, Producing Salvia. Cooperative Extension Service, Michigan State University, Extension Bulletin E-1663.
• Karam, N. S., Jawad, F. M., Arıkat, N. A. ve Shibli, R A. (2003). Growth and rosmarinic acid accumulation in callus, cell suspension, and root cultures of wild Salvia fruticosa. Plant Cell, Tissue and Organ Culture, 73, 117-121. https://doi.org/10.1023/A:1022806420209
• Kocabaş, I., Sönmez, İ., Kalkan, H. ve Kaplan, M. (2007). Farklı organik gübrelerin adaçayı (Salvia fruticosa Mill.)’nın uçucu yağ oranı ve bitki besin maddeleri içeriğine etkileri. Akdeniz Üniv. Ziraat Fak. Dergisi, 20(1), 105-110. https://dergipark.org.tr/tr/download/article-file/18136
• Makunga, N. P. ve van Staden, J. (2008). An efficient system forthe production of clonal plantlets of the medicinally important aromatic plant: Salvia africana-lutea L. Plant Cell Tiss Organ Cult, 9, 63-72. https://doi.org/10.1007/s11240-007-9305-5
• Murashige, T. ve Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiology Plantarum, 15, 473-497. https://florestal81.webnode.com/_files/200000040-03153040fe/07%20Artigo%20MS%201962.pdf
• Musarurwa, H. T., van Staden, J. ve Makunga, N. P. (2010). In vitro seed germination and cultivation of the aromatic medicinal Salvia stenophylla (Burch. ex Benth.) provides an alternative source of α-bisabolol. Plant Growth Regul., 61, 287-295. https://doi.org/10.1007/s10725-010-9476-7
• Özcan, İ.İ., Arabacı, O. ve Öğretmen, N.G. (2014). Bazı adaçayı türlerinde farklı tohum çimlendirme uygulamalarının belirlenmesi. Türk Tarım-Gıda Bilim ve Teknoloji Dergisi, 2(5), 203-207. http://www.agrifoodscience.com/index.php/TURJAF/article/view/58/52
• Petrova, M., Nikolova, M., Dimitrova, L. ve Zayova, E. (2015). Micropropagation and evaluation of flavonoid content and antioxidant activity of Salvia officinalis L. Genetics and Plant Physiology, 5(1), 48-60. https://www.researchgate.net/publication/301888959
• Santos-Gomes, P.C., Seabra, M.R., Andrade, P.B. ve Fernandes-Ferreira, M. (2002). Phenolic antioxidant compounds produced by in vitro shoots of sage (Salvia officinalis L.). Plant Science, 162, 981-987. https://doi.org/10.1016/S0168-9452(02)00052-3
• Skała, E. ve Wysokińska, H. (2004). In vitro regeneration of Salvia nemorosa L. from shoot tips and leaf explants. In Vitro Cell Dev. Biol.-Plant, 40, 596-602. https://doi.org/10.1079/IVP2004580
• Soundararajan, P., Sivanesan, I., Jo, E. H. ve Jeong, B.R. (2013). Silicon promotes shoot proliferation and shoot growth of Salvia splendens under salt stress in vitro. Hort. Environ. Biotechnol., 54(4), 311-318. https://doi.org/10.1007/s13580-013-0118-7
• Surgun-Acar, Y. ve Bürün, B. (2017, May, 9-11). Plant tissue culture studies in genus Salvia and its importance. International Congress on Medicinal and Aromatic Plants:“Natural and Healthy Life” (TABKON17), 09-11 May 2017, Proceedings Book, p.303-309, Konya, Türkiye.
• Surgun-Acar, Y., Yokaş, İ. ve Bürün, B. (2017, July, 5-8). Seed germination studies in native species for conservation of biodiversity. The 3rd International Symposium on EuroAsian Biodiversity (SEAB 2017), Abstract e-Book page:663, Minsk, Belarus.
• Thanos, C.A ve Doussi, M.A. (1995). Ecophysiology of seed germination in endemic labiates of crete. Israel Journal of Plant Sciences, 43, 227-237. https://doi.org/10.1080/07929978.1995.10676607
• Wu, Q., Zhang, C., Yang, H., Sun, Y., Hu, J. ve Zou, L., (2022). In vitro propagation via organogenesis and formation of globular bodies of Salvia plebeia: A valuable medicinal plant. In Vitro Cellular & Developmental Biology – Plant, 58, 51–60. https://doi.org/10.1007/s11627-021-10223-y
• Yılmaz-Gökdoğan, E. ve Bürün, B. (2021, July, 1-3). The seed germination and in vitro culture studies on Salvia species in flora of Turkey. The 5th International Symposium on EuroAsian Biodiversity (SEAB2021), Muğla, Turkey.
• Zayova, E., Nikolova, M., Dimitrova, L. ve Petrova, M. (2016). Comparative study of in vitro, ex vitro and in vivo propagated Salvia hispanica (Chia) plants: Morphometric analysis and antioxidant activity. AgroLife Scientific Journal, 5(2), 166-173. http://agrolifejournal.usamv.ro/pdf/vol.V_2/Art27.pdf